Original Research

Heart Rate Variability After Sprint Interval Training

in Cyclists and Implications for Assessing
Physical Fatigue
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Rafał G. Hebisz, Paulina Hebisz, and Marek W. Zatoń

Department of Physiology and Biochemistry, University School of Physical Education, Wroclaw, Poland

Abstract
Hebisz, RG, Hebisz, P, and Zatoń, MW. Heart rate variability after sprint interval training in cyclists and implications for assessing
physical fatigue. J Strength Cond Res 36(2): 558–564, 2022—This study evaluated the time- and frequency-domain indexes of
heart rate variability (HRV) during sprint interval exercise test (SIXT) and identify the onset of fatigue by HRV concurrent with changes
in average (Pavg) and peak (Ppeak) power output, total oxygen uptake (V̇O2tou), and blood hydrogen (H+) and lactate (La2)
concentrations. Twenty-seven cyclists performed 4 sets of SIXT in which each set consisted of four 30-second maximal sprints
interspersed with 90 seconds of low-intensity cycling. Each set was separated by 25–40 minutes of recovery. Before beginning
each set, HRV was analyzed by time (mean normal-to-normal RR intervals [RRNN], SD of normal-to-normal RR intervals [SDNN],
and square root of the mean squared difference between successive normal-to-normal RR intervals [RMSSD]) and frequency (total
spectral power [T] and very low- [VLF], low- [LF], and high-frequency [HF] spectral power) domain methods. Pavg, Ppeak, and
V̇O2tou were recorded in each set, and H+ and La2 were measured after each set. RRNN, SDNN, and VLF decreased in the second
set, whereas all time and frequency indexes of HRV decreased in the third and fourth set. Pavg and H+ decreased, while V̇O2tou
increased in the fourth set. Ppeak decreased in the second, third, and fourth set. Correlations were found between the changes in
the time and frequency indexes of HRV with H+, La2, and V̇O2tou. The results indicate that HRV does not reflect the onset of physical
fatigue in SIXT as was observed in Pavg and no correlation was found between the changes in HRV with Pavg and Ppeak.
Key Words: fatigue, maximal sprints, the time and frequency domain indexes of HRV, power output

Introduction and exercise performance and in monitoring training-induced fa-

Heart rate (HR) is modulated by the balance of the para-
sympathetic and the sympathetic divisions of the autonomic
nervous system (ANS). The increase in HR that occurs from rest
to lower-intensity exercise is first attributed to a decrease in
parasympathetic activity known as vagal withdrawal. As exercise
intensity increases, various neural mechanisms increase sympa-
thetic tone to induce a further rise in HR (40). Other stimuli such
as emotional arousal can modulate HR, in which anger or mental
stress can increase sympathetic activity and therefore HR (30). A
popular and noninvasive measure of ANS function and its re-
sponse to various stressors is the variation in the time interval
between heartbeats (RR) otherwise known as HR variability
(HRV). This measure and several related indexes are considered
to reflect changes in autonomic regulation (primarily para-
sympathetic nervous system activity) and can be analyzed by
time- and frequency-domain methods (3).
Although HRV is typically evaluated during rest, recent research
suggests that measuring HRV kinetics in response to exercise stres-
sors may have considerable potential as a predictor of aerobic fitness
Address correspondence to Dr. Paulina Hebisz, paulinahebisz@interia.pl.
Journal of Strength and Conditioning Research 36(2)/558–564
Copyright ª 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf
of the National Strength and Conditioning Association. This is an open access article
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No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and
share the work provided it is properly cited. The work cannot be changed in any way
or used commercially without permission from the journal.
tigue of elite athlete (41,42). Heart rate variability assessed after the
cessation of exercise has also recently been used as a marker of
parasympathetic reactivation to indicate postexercise recovery status
(33) and as an indicator of training load (exercise intensity and time)
(23,24). After acute exercise, time-domain measures such as mean
normal-to-normal RR intervals (RRNN) and the SD of normal-to-
normal RR intervals (SDNN) are lower than pre-exercise values.
These changes are accompanied with significantly modulated HRV
frequency-domain measures, from very low-frequency spectral
power (VLF) to high-frequency spectral power (HF), and have been
observed in both untrained (37) and trained (7) individuals. Michael
et al. (33) indicated that low-frequency spectral power (LF) is de-
pendent on both sympathetic and parasympathetic ANS activity and
that the ratio of low-frequency to high-frequency spectral power (LF/
HF) is indicative of sympathovagal balance where one system
dominates over the other. Changes in the above HRV frequency-
domain parameters are greater after longer and more intense efforts
and indicate a reduction in parasympathetic nervous system activity
(25,43). The recovery of parasympathetic system activity is believed
to be dependent primarily on an exercise intensity-dependent com-
bination of metabolite accumulation in blood and skeletal muscle as
well as changes in blood plasma and resultant arterial-baroreflex
stimulation (43). The postexercise reactivation of parasympathetic
system activity relative to exercise intensity has been observed in
studies involving both athletes (9) and inactive cohorts (44). In the
former, Bucheit et al. (9) observed an increased delay in para-
sympathetic recovery after a series of maximal all-out sprints

558
Heart Rate Variability and Physical Fatigue (2022) 36:2
compared with moderate-intensity continuous running. In the latter,
Stuckey et al. (44) reported reduced parasympathetic reactivation
after 4 repeated sprint intervals when compared with a single sprint
interval, although total recovery time in both protocols was above 60
minutes. Several authors have accredited the prolonged recovery of
parasympathetic activity (primarily vagal reactivation) after higher-
intensity exercise because of the greater anaerobic energy contribu-
tion in these types of efforts (9,29).
Research on parasympathetic nervous system reactivation after
exercise has focused primarily on the postexercise period after
a single dose of high-intensity interval training exercise (9,24,44).
Less is known about ANS modulation when assessed by the time-
and frequency-domain indexes of HRV particularly after sprint in-
terval training, in which maximal sprints are repeated in sets sepa-
rated by low- or moderate-intensity exercise of an extended
duration. Investigating the effects of such maximal efforts can en-
hance knowledge on the time course of parasympathetic nervous
system reactivation and correlate such data with training duration
(such as the number of completed sprint repetitions). In addition to
using HRV as an indicator of training load (23,24), several authors
have suggested that HRV indexes can be used to assess fatigue status
(14,28,38). However, these studies treated fatigue as accumulated
physical fatigue induced by overreaching or overtraining during
periods of several months, and in 1 case, the occurrence of a fatigue
state was not accompanied by a decrease in total power output in a 5-
minute maximal cycling test (38). Furthermore, the remaining
studies either did not analyze pre-training and post-training differ-
ences in power and work output or quantified fatigue only by sub-
jective ratings of perceived exertion which have certain limitations
(17,28,41). For these reasons, research on the effects of repeated
maximal sprints (4 sets of the sprint interval exercise test [SIXT]) on
HRV recovery dynamics and correlations with various physiological
measures of fatigue, including changes in power output, can provide
a springboard of important data. It is possible that HRV can be used
to predict power output before the execution maximal sprint and
therefore serve as an important indicator of future exercise perfor-
mance (12). More recent studies have suggested that baseline cardiac
autonomic function, determined on the basis of LF and HF spectral
analysis and its acute response to all-out interval exercise, can elu-
cidate the individual physiological responses (workload and peak
oxygen uptake) to high-intensity interval training (26).
The aim of this study was to therefore evaluate the time- and
frequency-domain indexes of HRV during a SIXT and identify the
onset of fatigue by HRV concurrent with changes in average and
peak power output, total oxygen uptake, and blood hydrogen and
lactate concentrations. Research involving repeated maximal sprints
is particularly warranted as it is a common training protocol in sports
that involve recurring bouts of high-intensity and low-intensity ex-
ercise such as in cross-country mountain biking (MTB). Depending
on the competitive MTB subdiscipline, uphill sections can last several
dozen seconds or even 20–30 minutes and the actual duration of the
race can be several minutes (cross-country eliminator races—XCE),
approximately 1 hour 30 minutes (cross-country Olympic races—
XCO), or even 4 hours 30 minutes (cross-country marathon
races—XCM) at the World Cup level. A SIXT that could emulate the
Table 1
Anthropometric and physiological characteristics.*†
Group Age (y) Height (cm)
22.3 6 6.1 178 6 7.4
*VȮ 2max 5 maximal oxygen uptake in the incremental exercise testing; Pmax 5 maximal aerobic power in the incremental exercise testing.
†Data are presented as mean 6 SD.
| www.nsca.com
structure of real-world MTB competition could provide practical
findings on repeated sprint performance and postexercise recovery
dynamics in this sporting domain.
It was hypothesized that HRV would decrease across when
performing repeated sets of maximal sprints and that HRV-
associated indexes would strongly correlate with the onset of
physical fatigue.
Methods
Experimental Approach to the Problem
The study would examine HRV indexes that correlate with the
onset of physical fatigue during a sets of repeated maximal sprints
in a SIXT. The use of HRV and HR data to predict fatigue could
allow the coach or conditioning specialist to prescribe an optimal
training stimulus by structuring training load to an individual’s
HRV changes compared with determining fatigue only after a de-
crease in power output, which induces the risk of overtraining.
Subjects performed 4 sets of SIXT in which each set consisted of
four 30-second maximal sprints interspersed with 90 seconds of low-
intensity cycling. Each set was separated by 25–40 minutes of re-
covery. A SIXT protocol was adopted to emulate cross-country
MTB competition where multiple maximal efforts (uphill sections)
are interspersed with low- and moderate-intensity cycling (downhill
sections) over a period of several hours. For the purposes of this
study, HRV indexes by time- and frequency-domain methods were
measured before beginning each set, average power output, peak
power output and total oxygen uptake during the test, and blood
hydrogen and lactate concentrations after completing each set.
Subjects
The study involved 27 well-trained MTB cyclists (21 males and 6
females). All had at least 3 years of competitive MTB experience with
yearly training time between 420 and 630 hours. Group anthropo-
metric characteristics and fitness level are presented in Table 1.
The study design was approved by the University School of
Physical Education in Wroclaw, Poland (chaired by Prof. Marek
Me˛ draś; on July 11, 2013) and performed in accordance with the
Declaration of Helsinki. All the cyclists were over 18 years of age
(age range: 18–34 years) and provided written informed consent
to participate in the study after being informed about the study’s
methods, procedures, benefits, and risks.
Procedures
The tests described below were performed in controlled labora-
tory conditions at the Exercise Laboratory at the University
School of Physical Education (PN-EN ISO 9001:2001 certified).
Subjects refrained from any training and strenuous physical ex-
ertion 48 hours before study outset. Subjects first completed an
incremental exercise test (IXT) to determine fitness level and after
48 hours of rest performed a SIXT for the purposes of the study.
Mass (kg) VȮ 2max (ml·kg21·min21) Pmax (W)
69.7 6 9.9 56.4 6 8.2 344.6 6 66.5
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Heart Rate Variability and Physical Fatigue (2022) 36:2
Testing was performed at the end of the preparatory season ap-
proximately 7–21 days before the beginning of the racing season.
Incremental Exercise Test. Testing was performed on a Cyclus 2
cycle ergometer (RBM elektronik-automation GmbH, Leipzig,
Germany) that was adjusted and calibrated before each trial.
Starting workload was set at 50 W and increased by another 50 W
every 3 minutes until volitional exhaustion was reached. If
a subject was unable to complete an entire 3-minute stage, 0.28 W
was subtracted for each missing second to determine individual
maximal aerobic power (Pmax). Respiratory gas exchange was
measured on a breath-by-breath basis with a Quark gas analyzer
(Cosmed, Milan, Italy). The device was calibrated before each test
with a reference gas mixture of carbon dioxide (5%), oxygen
(16%), and nitrogen (79%). Tidal air was collected by wearing
a face mask and analyzed to determine oxygen uptake (V̇ O2),
carbon dioxide excretion (V̇ CO2), and minute pulmonary venti-
lation (V̇ E). Absolute and relative (per kg of body mass) V̇ O2max
was calculated based on the composition of expired air and
minute ventilation. All measures were averaged over 30-second
intervals.
Sprint Interval Exercise Test. This test was performed on the same
Cyclus 2 cycle ergometer. Braking resistance was set to an in-
dividual fixed torque of 0.8 Nm·kg21. The test was preceded by
a 20-minute warm-up during which the subject cycled for 5
minutes at an intensity corresponding to 40% Pmax (as de-
termined in the IXT) and then for 15 minutes at 50% Pmax. An
active 10-minute cool-down was then performed by cycling at
10% Pmax followed by 5 minutes of passive recovery spent sitting
on the ergometer with arms resting on the handlebars. The test
proper involved 4 sets, with each set consisting of four 30-second
maximal sprints interspersed with 90 seconds of low-intensity
cycling. The each set was separated by 25–40 minutes of recovery
in which (a) the first 2 minutes involved low-intensity cycling; (b)
then, moderate-intensity cycling at 50% Pmax was continued until
blood pH returned to at least 7.35, resulting in individual re-
covery times from 18 to 33 minutes (blood pH was continually
Figure 1. Illustration of the sprint interval exercise test protocol concomitant with subject heart
rate (HR, heart rate, V̇O2tou 5 total oxygen uptake, Pavg 5 average power output, Ppeak 5
peak power output, H+ 5 blood hydrogen concentrations, La2 5 blood lactate concen-
trations, HRV 5 heart rate variability).
560
measured with a RAPIDLab 348 blood gas system—Siemens
Healthcare, Germany); and (c) at the end of the recovery, the
subject sat passively on the ergometer with arms resting on the
handlebars for 5 minutes before commencing another SIXT set.
Heart rate was continually monitored, and a visual representation
of the SIXT protocol across the maximal sprints and recovery is
presented in Figure 1.
RR intervals were recorded in the last 2 minutes of the
5-minute passive recovery when the subject was sitting on the
ergometer just before beginning another set of sprints. A V800
cardiofrequencimeter (Polar, Oy, Finland) integrated with Kubios
HRV Standard software (KubiosOy, Kuopio, Finland) was used
to analyze HRV by time and frequency domains. Time-domain
measures included the mean normal-to-normal RR intervals
(RRNN), SD of normal-to-normal RR intervals (SDNN), and the
square root of the mean squared difference between successive
normal-to-normal RR intervals (RMSSD). For the frequency
domain, spectral analysis was performed using Fast Fourier
Transformation to obtain very low-frequency spectral power
(VLF; 0.003–0.05 Hz), low-frequency spectral power (LF;
0.05–0.15 Hz), high-frequency spectral power (HF; 0.15–0.4
Hz), total spectral power (T), and the LF/HF ratio as a measure of
sympathovagal balance.
Power output (relative to body mass through individualized
braking resistance) was continually recorded by the ergometer
and used to calculate average power output (Pavg) and peak
power output (Ppeak) for each set. Respiratory function was
evaluated using the same procedures and equipment as in the IXT.
Gas exchange was measured from the beginning of each set until 2
minutes after set termination. Total oxygen uptake for each set
(V̇ O2tou) was determined as the amount of oxygen uptake across
the four 30-second sprints and 90-second recovery components.
Arterialized capillary blood was collected by squeezing the finger
to draw blood into a capillary tube 3 minutes after the fourth
sprint in each set to determine hydrogen (H1) and lactate (La2)
concentrations with a RAPIDLab 348 blood gas analyzer (Sie-
mens Healthcare, Erlangen, Germany) and LP400 photometer
(Dr. Lange, Burladingen, Germany), respectively.
Heart Rate Variability and Physical Fatigue (2022) 36:2 | www.nsca.com

Statistical Analyses fourth sets compared with the first set. In addition, RRNN,
SDNN, and VLF had significantly decreased in the second set
The data set was analyzed with the Statistica 13.1 software
compared with the first set. No significant differences were ob-
package (Statsoft). Arithmetic mean values and SDs were calcu-
served for LF/HF (F 5 1.32; p 5 0.275) (Table 2).
lated for all measures. Data were screened for outliers with the
Comparisons of the physiological and biochemical measures
three-sigma method, and all measures (3 subjects) outside of 3
showed significant differences for V̇ O2tou (F 5 4.88; p 5 0.004),
SDs of the mean were removed. The normality of the data dis-
H1 (F 5 10.06; p 5 0.000), Ppeak (F 5 8.86; p 5 0.000), and
tribution was then examined with the Shapiro-Wilk test. One-
Pavg (F 5 8.33; p 5 0.000) across the sets. Post hoc analysis
way repeated-measures analysis of variance (ANOVA) with
revealed the following significant changes: V̇ O2tou increased in
Scheffe’s test post hoc was used to compare the means across each
the fourth set compared with the first set, H1 decreased in the
of the 4 sets. To estimate the magnitude of effect sizes, the eta-
fourth set compared with the first and second sets, Ppeak de-
square (h2) was calculated for 1-way ANOVA with repeated
creased in the second, third, and fourth sets compared with the
measurements.
first set, and Pavg decreased in the fourth set compared with the
Correlation analysis was performed by first calculating the
second set. No significant differences were found for La2 (F 5
absolute difference for each variables between the SIXT sets
1.70; p 5 0.176) (Table 3).
for each subject. The differences between the first and second
Correlative analysis showed that dV̇ O2tou weakly correlated
set, first and third set, and first and fourth set were de-
with dSDNN, dVLF, and dT and moderately with dRRNN. In
termined to obtain 72 values in total (3 values 3 24 subjects).
turn, dH1 was moderately correlated with dRRNN, dSDNN,
These differences (d) were tabulated for each variable
and dRMSSD, whereas dLa2 weakly correlated with dSDNN and
(dRRNN, dSDNN, dRMSSD, dVLF, dLF, dHF, dT, dLF/HF,
moderately with dRMSSD. No significant correlations were ob-
dPavg, dPpeak, dV̇ O2 tou, dH 1 , and dLa 2 ), and Pearson’s
served for dPpeak or dPavg with any of the between-set differ-
correlation coefficients (r) were determined between the dif-
ences in HRV or between dH1, dLa2, and any of the between-set
ferences in the HRV indexes (dRRNN, dSDNN, dRMSSD,
differences for any of the frequency-domain indexes of HRV
dVLF, dLF, dHF, dT, and dLF/HF) and physiological and
(Table 4).
biochemical measures (dV̇ O 2 tou, dH1 , dLa 2 , dPpeak, and
dPavg). The level of significance was set at 0.05 for all
procedures.
Discussion
The present results showed a positive correlation between the
Results
changes in the time-domain indexes of HRV with H 1 and La 2
The 1-way repeated-measures ANOVA indicated significant dif- concentrations yet a negative correlation between the
ferences for RRNN (F 5 37.57; p 5 0.000), SDNN (F 5 20.95; changes in the time-domain (RRNN and SDNN) and
p 5 0.000), RMSSD (F 5 7.36; p 5 0.000), VLF (F 5 8.56; p 5 frequency-domain (VLF and T) indexes of HRV with V̇ O 2tou.
0.000), LF (F 5 9.86; p 5 0.000), HF (F 5 5.61; p 5 0.002), and T Between-set comparisons showed increasingly greater
(F 5 10.37; p 5 0.000) across the sets. Post hoc analysis showed reductions in SDNN, RMSSD, and HF with repeated maxi-
significant decreases in all of the above variables in the third and mal sprints (quantified as the number of completed SIXT

Table 2
Heart rate variability indexes during the last 2 minutes of passive recovery component before each sprint interval exercise test set.*†
Pre-1st set Pre-2nd set Pre-3rd set Pre-4th set ES
Time domain (ms)
RRNN 544.7 6 67.7 492.9 6 49.6ठ473.4 6 36.6ठ471.2 6 37.4ठ0.62
95% CI: U│L 573.3 │ 516.1 513.8 │ 472.0 488.8 │ 457.9 486.9 │ 455.4
SDNN 20.6 6 11.0 12.6 6 8.3ठ8.9 6 5.5ठ8.2 6 4.8ठ0.48
95% CI: U│L 25.2 │ 15.9 16.1 │ 9.1 11.3 │ 6.6 10.2 │ 6.1
RMSSD 13.9 6 14.0 7.6 6 6.1 5.2 6 4.1ठ4.6 6 2.6ठ0.24
95% CI: U│L 19.9 │ 8.0 10.2 │ 5.0 6.9 │ 3.5 5.7 │ 3.5
Frequency domain (ms2)
VLF 39.5 6 43.7 18.6 6 20.6ठ11.5 6 12.7ठ10.7 6 15.0ठ0.27
95% CI: U│L 57.9 │ 21.0 27.3 │ 9.9 16.9 │ 6.1 17.0 │ 4.4
LF 266.6 6 289.7 187.0 6 301.6 65.0 6 92.0‡§ 55.9 6 64.2‡§‖ 0.30
95% CI: U│L 388.9 │ 144.3 314.4 │ 59.6 103.8 │ 26.2 83.0 │ 28.8
HF 53.5 6 68.8 34.0 6 91.6 12.8 6 23.5ठ7.6 6 10.1ठ0.20
95% CI: U│L 82.6 │ 24.5 72.7 │ 24.6 22.7 │ 2.9 11.9 │ 3.4
T 359.8 6 366.7 239.7 6 407.1 89.2 6 123.4ठ74.2 6 76.6ठ0.31
95% CI: U│L 514.6 │ 204.9 411.7 │ 67.8 141.3 │ 37.1 106.5 │ 41.9
LF/HF 6.3 6 3.4 9.4 6 7.1 8.4 6 6.7 8.8 6 6.7 0.05
95% CI: U│L 8.0 │ 4.7 12.4 │ 6.3 11.2 │ 5.5 11.7 │ 6.0
*RRNN 5 mean normal-to-normal RR intervals; SDNN 5 SD of normal-to-normal RR intervals; RMSSD 5 square root of the mean squared difference between successive normal-to-normal RR intervals; VLF
5 very low-frequency spectral power; LF 5 low-frequency spectral power; HF 5 high-frequency spectral power; T 5 total spectral power; LF/HF 5 ratio of low-frequency to high-frequency spectral power.
†Data are presented as mean 6 SD with effect sizes (ES) and 95% confidence intervals: upper│lower (95% CI: U│L).
‡p , 0.05.
§Significantly different from the first set.
‖Significantly different from the second set.

561
Heart Rate Variability and Physical Fatigue (2022) 36:2

Table 3
Physical, physiological, and biochemical measures in each sprint interval exercise test set.*†
1st set 2nd set 3rd set 4th set ES
Ppeak [W] 1,253.7 6 315.0 1,154.4 6 294.6‡,§ 1,146.8 6 302.4‡,§ 1,139.9 6 295.4‡,§ 0.28
95% CI: U│L 1,386.7 │ 1,120.6 1,278.8 │ 1,030.0 1,274.5 │ 1,019.1 1,264.6 │ 1,015.1
Pavg [W] 604.0 6 120.8 612.2 6 120.3 604.8 6 118.7 596.6 6 117.1‡,‖ 0.27
95% CI: U│L 655.0 │ 552.9 663.0 │ 561.4 654.9 │ 554.7 643.1 │ 544.1
VȮ 2tou [L] 19.06 6 3.94 19.41 6 4.11 19.70 6 4.01 20.11 6 4.24‡,§ 0.17
95% CI: U│L 20.73 │ 17.40 21.14 │ 17.67 21.39 │ 18.01 21.89 │ 18.31
H1 [nmol·L21] 89.32 6 9.07 85.71 6 8.33 85.21 6 9.06 81.42 6 11.06‡,§,‖ 0.30
95% CI: U│L 93.16 │ 85.48 89.24 │ 82.24 89.04 │81.42 86.19 │76.84
La-[mmol·L21] 17.70 6 2.33 17.89 6 1.85 17.83 6 1.87 17.06 6 2.53 0.07
95% CI: U│L 18.68 │ 16.72 18.67 │ 17.11 18.62 │ 17.04 18.13 │ 15.99
*Ppeak 5 peak power output; Pavg 5 average power output; VȮ 2tou 5 total oxygen uptake; H1 5 blood hydrogen concentration; La2 5 blood lactate concentration.
‡p , 0.05.
§Significantly different from the first set.
‖Significantly different from the second set.

sets), suggesting reduced parasympathetic nervous system phosphocreatine (10), the onset of metabolic acidosis (36), and
activity, primarily in vagal tone, and/or prolonged para- the excess accumulation of ADP and phosphate (2,8). How-
sympathetic reactivation in trained MTB cyclists. These ever, Fitts (16) and Westerblad et al. (45) suggest that low pH
findings support the notion that total exercise time is a de- resulting from high concentrations of H1 is the principal factor
terminant of postexercise parasympathetic recovery and responsible for muscle fatigue. We observed H1 concen-
HRV-related measures (5,21,23,24,44). Interestingly, the trations exceeding 80 nmol·L21 in all 4 sets, far above normal
changes in the time- and frequency-domain indexes of HRV resting values (38–44 nmol·L21) and indicative of metabolic
registered in the second, third, and fourth sets were signifi- acidosis (15,22,39). As had been indicated by other research-
cant only in relation to the first set. No significant changes in ers, the accumulation of H1 can reduce parasympathetic ner-
the indexes of HRV were observed when comparing the sec- vous system activity (13,43) and that parasympathetic
ond set with the third and fourth sets (excluding LF) or when reactivation can exceed 1 hour after sprint interval training
comparing the third set with the fourth set, suggesting a finite protocol similar to one used in this study (44). In our study, H1
limit of HRV kinetics after repeated maximal sprints and decreased only after the fourth set with no significant changes
high-intensity exercise (23,24). in La2 across successive sets in the SIXT, suggesting that the
Previous studies have posited that the changes observed in changes in the time-domain indexes of HRV may depend on
the time- and frequency-domain indexes of HRV can be used to several factors independent of exercise intensity and the post-
assess fatigue status in trained subjects (41). Although early exercise accumulation of metabolites in blood and skeletal
research attributed fatigue exclusively to the accumulation muscle. One possible explanation is the increase in blood
of H1 (19), later findings have suggested several cause-and- plasma volume with additional sets of maximal sprints (43),
effect models (1,4,35). These include the inability of the which was observed in a previous study that applied an iden-
heart to deliver sufficient oxygenated blood to working tical SIXT protocol (20).
muscle including cardiomyocytes resulting in a depletion of The literature defines fatigue as the inability to maintain
a prescribed exercise intensity or a reduction in power output
(6,34), with fatigue in short-duration sprints (,10 seconds)
quantified as a decrease in both peak power and mean power
Table 4
(18,32). We observed a reduction in Ppeak in the second, third,
Pearson’s correlations coefficients (r) for the between-set
and fourth sets, whereas Pavg decreased only in the fourth set.
differences of heart rate variability indexes and physiological,
biochemical, and physical measures.*
Although this study involved 30-second and not 10-second
sprints, the applied exercise protocol appears to be insufficient
dVȮ 2tou dH1 dLa2 dPpeak dPavg
in significantly decreasing average power output. This finding
dRRNN 20.40† 0.33† 0.03 0.04 0.02 challenges the prognostic significance of the time-domain in-
dSDNN 20.27† 0.35† 0.29† 20.04 0.03
dexes of HRV in the assessment of fatigue during repeated
dRMSSD 20.17 0.36† 0.43† 20.06 0.02
dVLF 20.25† 20.01 20.09 20.11 20.23
maximal sprints particularly in well-trained athletes. It is
dLF 20.23 0.20 0.08 0.10 20.09 possible that the active recovery component between each set
dHF 20.22 20.03 0.21 20.09 20.09 was sufficient in duration to eliminate the onset of peripheral
dT 20.26† 0.16 0.09 0.04 20.12 fatigue (8,11). Another reason may lie with the high aerobic
dLF/dHF 0.08 0.02 20.23 0.03 0.06 fitness level of the recruited subjects. Girard et al. (18) reported
that trained athletes do not show large decreases in power
*d suffix 5 difference between values obtained in the first and the second set, first and the third set,
and first and the fourth set calculated for each subject and them for the entire sample; RRNN 5 mean output during repeated sprint (,10 seconds) exercise, whereas
normal-to-normal RR intervals; SDNN 5 SD of normal-to-normal RR intervals; RMSSD 5 square root Menaspa et al. (31) showed that elite cyclists generate similar
of the mean squared difference between successive normal-to-normal RR intervals; VLF 5 very low- levels of power output during repeated sprint exercise re-
frequency spectral power; LF 5 low-frequency spectral power; HF 5 high-frequency spectral power; gardless of the preceding mode of exercise (10 minutes of
T 5 total spectral power; LF/HF 5 ratio of low-frequency to high-frequency spectral power; VȮ 2tou,
total oxygen uptake; H1 5 blood hydrogen concentration; La2 5 blood lactate concentration; Ppeak
nonvariable cycling at 93% HRmax vs. 10 minutes variable
5 peak power output; Pavg 5 average power output. cycling at an intensity approximately 93% HRmax). Other
†p , 0.05. factors that may have modulated the onset of fatigue in certain

562
Heart Rate Variability and Physical Fatigue (2022) 36:2
individuals external to a disrupted acid-base balance including
sodium-potassium pump inhibition and a decrease in mem-
brane excitability reduced motor unit recruitment and firing
rate (18) and a greater depletion of muscle glycogen stores (1).
Nonetheless, the above results coupled with the lack of cor-
relation between Ppeak or Pavg and the between-set differ-
ences in any of the HRV indexes might indicate that the
development of fatigue in this scenario cannot be measured
by HRV.
As previously mentioned, relatable research treated fatigue
as overreaching during the training macrocycle and that only
some studies analyzed changes in performance measures
(17,41) or, those that did, reported no change in power output
with regard to changes in HRV (38). Only 1 study in the lit-
erature observed a reduction in IXT-obtained maximal power
during an overload training period concurrent with changes in
HRV, although these measures were averaged across a 7-day
period and therefore not comparable with acute pre-exercise or
post-exercise values (27). As a result, there is limited evidence
supporting the relationship between HRV and fatigue other
than fatigue induced by a significant acid-base disturbance
such as after exercise with a large anaerobic glycolytic com-
ponent. Hence, it may be that changes in HRV are more likely
the result of significant work performed and not indicative of
the onset of physical fatigue after repeated sets of maximal
sprints (9,24,27,44).
Practical Applications
Changes in the time- and frequency-domain indexes of
HRV do not reflect the onset of physical fatigue in SIXT, as
in 3 subsequent sets, no change in Pavg was observed and
no correlation was found between the changes in the time-
and frequency-domain indexes of HRV with Ppeak or
Pavg.
Based on our results, the development of fatigue before a set
of maximal sprints cannot be predicted by observing HRV
during SIXT in trained cyclists.
Acknowledgments
Supported by the Ministry of Science and Higher Education
under a Grant number NRSA300253. The program: Develop-
ment of Academic Sport.
The authors thank Michael Antkowiak for his translation of the
manuscript and language assistance.
The authors have no conflicts of interest to disclose.
References
1. Abbiss CR, Laursen PB. Models to explain fatigue prolonged endurance
cycling Turing. Sports Med 35: 865–898, 2005.
2. Ament W, Verkerke G. Exercise and fatigue. Sports Med 39: 389–422,
2009.
3. Aubert AE, Seps B, Beckers F. Heart rate variability in athletes. Sports Med
33: 889–919, 2003.
4. Bangsbo J, Graham TE, Kiens B, Saltin B. Elevated muscle glycogen and
anaerobic energy production during exhaustive exercise in man. J Physiol
451: 205–227, 1992.
5. Barak OF, Ovcin ZB, Jakovljevic DG, et al. Heart rate recovery after
submaximal exercise in four different recovery protocols in male athletes
and non-athletes. J Sports Sci Med 10: 369–375, 2011.
6. Black MI, Jones AM, Blackwell JR, et al. Muscle metabolic and neuro-
muscular determinants of fatigue during cycling in different exercise in-
tensity domains. J Appl Physiol 122: 446–459, 2017.
563
| www.nsca.com
7. Brown SJ, Brown JA. Resting and post exercise cardiac autonomic control
in trained master athletes. J Physiol Sci 57: 23–29, 2007.
8. Broxterman RM, Layec G, Hureau TJ, Amann M, Richardson RS. Skel-
etal muscle bioenergetics during all-out exercise: Mechanistic insight into
the oxygen uptake slow component and neuromuscular fatigue. J Appl
Physiol 122: 1208–1217, 2017.
9. Buchheit M, Laursen PB, Ahmaidi S. Parasympathetic reactivation after
repeated sprint exercise. Am J Physiol Heart Circ Physiol 293: 133–141,
2007.
10. Burnley M, Vanhatalo A, Fulford J, Jones AM. Similar metabolic per-
turbations during all-out and constant force exhaustive exercise in
humans: a31P magnetic resonance spectroscopy study. Exp Physiol 95:
798–807, 2010.
11. Cairns SP. Lactic acid and exercise performance. Culprit or friend? Sports
Med 36: 279–291, 2006.
12. Chalencon S, Pichot V, Roche F, et al. Modeling of performance and ANS
activity for predicting future responses to training. Eur J Appl Physiol 115:
589–596, 2015.
13. De Oliveira TP, Mattos RA, Da Silva RBF, Rezende RA, De Lima JRP.
Absence of parasympathetic reactivation after maximal exercise. Clin
Physiol Funct Imaging 33: 143–149, 2013.
14. Earnest CP, Jurca R, Church TS, et al. Relation between physical exertion
and heart rate variability characteristics in professional cyclists during
Tour of Spain. Br J Sports Med 38: 568–575, 2004.
15. Fitts RH. The cross-bridge cycle and skeletal muscle fatigue. J Appl
Physiol 104: 551–558, 2008.
16. Fitts RH. The role of acidosis in fatigue: Pro perspective. Med Sci Sports
Exerc 48: 2335–2338, 2016.
17. Flatt AA, Hornikel B, Esco MR. Heart rate variability and psychometric
responses to overload and tapering in collegiate sprint-swimmers. J Sci
Med Sport 20: 606–610, 2017.
18. Girard O, Mendez-Villanueva A, Bishop D. Repeated-sprint
ability—part I. Factors contributing to fatigue. Sports Med 41:
673–694, 2011.
19. Hartree W, Hill AV. The anaerobic processes involved in muscular ac-
tivity. J Physiol 58: 127–137, 1923.
20. Hebisz P, Hebisz R, Bakońska-Pacoń E, Zatoń M. Acute hematological
response to a single dose of sprint interval training in competitive cyclists.
Sci Sports 32: 369–375, 2017.
21. Hebisz P, Hebisz R, Zatoń M, Borkowski J. Dynamics of changes in
power output, heart rate, and disorders of acid-base balance during
interval training in mountain cyclists. Isokinet Exerc Sci 23: 245–252,
2015.
22. Hermansen L, Osnes JB. Blood and muscle pH after maximal exercise in
man. J Appl Physiol 32: 304–308, 1972.
23. Kaikkonen P, Hynynen E, Mann T, Rusko H, Nummela A. Can HRV be
used to evaluate training load in constant load exercises? Eur J Appl
Physiol 108: 435–442, 2010.
24. Kaikkonen P, Hynynen E, Mann T, Rusko H, Nummela A. Heart rate
variability is related to training load variables in interval running exer-
cises. Eur J Appl Physiol 112: 829–838, 2012.
25. Kaikkonen P, Rusko H, Martinmaki K. Post-exercise heart rate variability
of endurance athletes after different high-intensity exercise interventions.
Scand J Med Sci Sports 18: 511–519, 2008.
26. Kiviniemi AM, Tulppo MP, Eskelinen JJ, et al. Autonomic function pre-
dicts fitness response to short-term high-intensity interval training. Int J
Sports Med 36: 915–921, 2015.
27. Le Meur Y, Pichon A, Schaal K, et al. Evidence of parasympathetic hy-
peractivity in functionally overreached athletes. Med Sci Sports Exerc 45:
2061–2071, 2013.
28. Leti T, Bricout VA. Interest of analyses of heart rate variability in the
prevention of fatigue states in senior runners. Auton Neurosci 173: 14–21,
2012.
29. Martinmaki K, Rusko H. Time-frequency analysis of heart rate variability
during immediate recovery from low and high intensity exercise. Eur J
Appl Physiol 102: 353–360, 2008.
30. McCraty R, Atkinson M, Tiller WA, Rein G, Watkins AD. The effects of
emotions on short-term power spectrum analysis of heart rate variability.
Am J Cardiol 76: 1089–1093, 1995.
31. Menaspa P, Martin DT, Victor J, Abbiss CR. Maximal sprint power in
road cyclists after variable and non variable high-intensity exercise.
J Strength Cond Res 29: 3156–3161, 2015.
32. Mendez-Villanueva A, Hamer P, Bishop D. Fatigue in repeated-sprint
exercise is related to muscle power factors and reduced neuromuscular
activity. Eur J Appl Physiol 103: 411–419, 2008.
Heart Rate Variability and Physical Fatigue (2022) 36:2
33. Michael S, Graham KS, Oam DGM. Cardiac autonomic responses during
exercise and post-exercise recovery using heart rate variability and systolic
time intervals—a review. Front Physiol 8: 301–319, 2017.
34. Millet GY, Lepers R. Alterations of neuromuscular function after pro-
longed running, cycling and skiing exercises. Sports Med 34: 105–116,
2004.
35. Nevill ME, Boobis LH, Brooks S, Williams C. Effect of training on muscle
metabolism during treadmill sprinting. J Appl Physiol 67: 2376–2382,
1989.
36. Noakes TD. Physiological models to understand exercise fatigue and the
adaptations that predictor enhance athletic performance. Scand J Med Sci
Sports 10: 123–145, 2000.
37. Parekh A, Lee CM. Heart rate variability after isocaloric exercise bouts of
different intensities. Med Sci Sports Exerc 37: 599–605, 2005.
38. Pichot V, Busso T, Roche F, et al. Autonomic adaptations to intensive and
overload training periods: A laboratory study. Med Sci Sports Exerc 34:
1660–1666, 2002.
564
39. Robergs RA, Ghiasvand F, Parker D. Biochemistry of exercise-induced
metabolic acidosis. Am J Physiol 287: R502–R516, 2004.
40. Robinson BF, Epstein SE, Beiser GD, Braunwald E. Control of heart rate
by the autonomic nervous system. Circ Res 19: 400–411, 1966.
41. Schmitt L, Regnard J, Desmarets M, et al. Fatigue shifts and scatters heart
rate variability in elite endurance athletes. PLoS One 8: 1–9, 2013.
42. Schmitt L, Regnard J, Parmentier AL, et al. Typology of “Fatigue” by
heart rate variability analysis in elite nordic-skiers. Int J Sports Med 36:
999–1007, 2015.
43. Stanley J, Peake JM, Buchheit M. Cardiac parasympathetic reactivation
following exercise: Implications for training prescription. Sports Med 43:
1259–1277, 2013.
44. Stuckey MI, Tordi N, Mourot L, et al. Autonomic recovery following
sprint interval exercise. Scand J Med Sci Sports 22: 756–763, 2012.
45. Westerblad H, Allen DG, L ännergren J. Muscle fatigue: Lactic acid or
inorganic phosphate the major cause? News Physiol Sci 17: 17–21,
2002.